基于抗原表位烟草环斑病毒单克隆抗体的制备  

Monoclonal antibody preparation of Tobacco ringspot virus based on antigenic epitopes

作　　者：秦绍钊[1,2] 何月秋[1] 李旻[2] 雷屈文[1,2] 王光勇[3] 丁元明[2] 

机构地区：[1]云南农业大学植物保护学院,昆明650201 [2]云南出入境检验检疫局技术中心,昆明650228 [3]昆明理工大学生命科学与技术学院,昆明650224

出　　处：《华中农业大学学报》2012年第5期598-603,共6页Journal of Huazhong Agricultural University

基　　金：国家质检总局科技计划项目(2010IK259)

摘　　要：基于多种生物信息学软件综合预测烟草环斑病毒(TRSV)外壳蛋白抗原表位所在的结构域,明确了该外壳蛋白抗原表位可能位于C端319～328、384～388、420～430、480～487、495～511位氨基酸残基附近。通过RT-PCR扩增出了目的片段,构建了TRSV外壳蛋白原核表达载体,表达的TRSV外壳蛋白外源融合蛋白具免疫活性。采用杂交瘤细胞技术,利用外源融合蛋白制备了单克隆抗体,制备的抗血清可与TRSV外壳蛋白基因原核表达产物发生免疫反应。检验结果表明,制备的抗血清具较好的特异性,可应用于TRSV的检疫检验。By using bioinformaties software, the domain of antigenic epitopes on Tobacco ringspot virus （TRSV） coat protein was analyzed and predicted, and the epitopes were found to be located near 319-328,384-388,420-430,480-487,495-511 amino acid residues at C-terminal. After the targeted frag- ment was amplified by RT-PCR based on the specific primers, the expression vector was constructed and the monoclonal antibody was prepared with the fusion protein and hybridoma. The antiserum prepared with the fusion protein reacted specifically with the expressed product of TRSV ccat protein in E. coli. The antiserum had high specificity and may be applied to quarantine and inspection.

关 键 词：烟草环斑病毒 抗原表位 原核表达 融合蛋白 单克隆抗体 

分 类 号：S41[农业科学—植物保护] S432.41